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Journal List > Biochem J > v.327(Pt 1); Oct 1, 1997

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Biochem J. 1997 October 1; 327(Pt 1): 305–310.
PMCID: PMC1218795
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ATP-dependent transport of bilirubin glucuronides by the multidrug resistance protein MRP1 and its hepatocyte canalicular isoform MRP2.
G Jedlitschky, I Leier, U Buchholz, J Hummel-Eisenbeiss, B Burchell, and D Keppler
Division of Tumor Biochemistry, Deutsches Krebsforschungszentrum, In Neuenheimer Feld 280, D-69120 Heidelberg, Federal Republic of Germany.
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Abstract
Bilirubin is secreted from the liver into bile mainly as monoglucuronosyl and bisglucuronosyl conjugates. We demonstrate for the first time that ATP-dependent transport of both bilirubin glucuronides is mediated by the multidrug resistance protein (MRP1) as well as by the distinct canalicular (apical) isoform MRP2, also termed cMRP or cMOAT (canalicular multispecific organic anion transporter). In membrane vesicles from MRP1-transfected HeLa cells mono[3H]glucuronosylbilirubin and bis[3H]glucuronosylbilirubin (each at 0.5 microM) were transported with rates of 5.3 and 3.1 pmol/min per mg of protein respectively. Rat hepatocyte canalicular membrane vesicles, which contain Mrp2 (the rat equivalent of MRP2), transported mono[3H]glucuronosylbilirubin and bis[3H]glucuronosylbilirubin at rates of 8.9 and 8.5 pmol/min per mg of protein, whereas membrane vesicles from mutant liver lacking Mrp2 showed no transport of the conjugates. In membrane vesicles from human hepatoma Hep G2 cells, which predominantly expressed MRP2, transport rates were 8.3 and 4.4 pmol/min per mg of protein for monoglucuronosylbilirubin and bisglucuronosylbilirubin respectively. ATP-dependent transport of the glutathione S-conjugate -3H-leukotriene C4, an established high-affinity substrate for MRP1 and MRP2, was inhibited by both bilirubin glucuronides with IC50 values between 0.10 and 0.75 microM. The ratios of leukotriene C4 transport and bilirubin glucuronide transport, determined in the same membrane vesicle preparation, indicated substrate specificity differences between MRP1 and MRP2 with a preference of MRP2 for the glucuronides.
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Selected References
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  • BILLING BH, COLE PG, LATHE GH. The excretion of bilirubin as a diglucuronide giving the direct van den Bergh reaction. Biochem J. 1957 Apr;65(4):774–784. [PubMed]
  • SCHMID R. The identification of direct-reacting bilirubin as bilirubin glucuronide. J Biol Chem. 1957 Dec;229(2):881–888. [PubMed]
  • Gordon ER, Goresky CA, Chang TH, Perlin AS. The isolation and characterization of bilirubin diglucuronide, the major bilirubin conjugate in dog and human bile. Biochem J. 1976 Jun 1;155(3):477–486. [PubMed]
  • Fevery J, Van de Vijver M, Michiels R, Heirwegh KP. Comparison in different species of biliary bilirubin-IX alpha conjugates with the activities of hepatic and renal bilirubin-IX alpha-uridine diphosphate glycosyltransferases. Biochem J. 1977 Jun 15;164(3):737–746. [PubMed]
  • Blanckaert N. Analysis of bilirubin and bilirubin mono- and di-conjugates. Determination of their relative amounts in biological samples. Biochem J. 1980 Jan 1;185(1):115–128. [PubMed]
  • Burchell B, Brierley CH, Rance D. Specificity of human UDP-glucuronosyltransferases and xenobiotic glucuronidation. Life Sci. 1995;57(20):1819–1831. [PubMed]
  • Kobayashi K, Sogame Y, Hayashi K, Nicotera P, Orrenius S. ATP stimulates the uptake of S-dinitrophenylglutathione by rat liver plasma membrane vesicles. FEBS Lett. 1988 Nov 21;240(1-2):55–58. [PubMed]
  • Ishikawa T, Müller M, Klünemann C, Schaub T, Keppler D. ATP-dependent primary active transport of cysteinyl leukotrienes across liver canalicular membrane. Role of the ATP-dependent transport system for glutathione S-conjugates. J Biol Chem. 1990 Nov 5;265(31):19279–19286. [PubMed]
  • Nishida T, Hardenbrook C, Gatmaitan Z, Arias IM. ATP-dependent organic anion transport system in normal and TR- rat liver canalicular membranes. Am J Physiol. 1992 Apr;262(4 Pt 1):G629–G635. [PubMed]
  • Keppler D, Kartenbeck J. The canalicular conjugate export pump encoded by the cmrp/cmoat gene. Prog Liver Dis. 1996;14:55–67. [PubMed]
  • Oude Elferink RP, Meijer DK, Kuipers F, Jansen PL, Groen AK, Groothuis GM. Hepatobiliary secretion of organic compounds; molecular mechanisms of membrane transport. Biochim Biophys Acta. 1995 Jul 17;1241(2):215–268. [PubMed]
  • Jansen PL, Peters WH, Lamers WH. Hereditary chronic conjugated hyperbilirubinemia in mutant rats caused by defective hepatic anion transport. Hepatology. 1985 Jul–Aug;5(4):573–579. [PubMed]
  • Jansen PL, Groothuis GM, Peters WH, Meijer DF. Selective hepatobiliary transport defect for organic anions and neutral steroids in mutant rats with hereditary-conjugated hyperbilirubinemia. Hepatology. 1987 Jan–Feb;7(1):71–76. [PubMed]
  • Kuipers F, Enserink M, Havinga R, van der Steen AB, Hardonk MJ, Fevery J, Vonk RJ. Separate transport systems for biliary secretion of sulfated and unsulfated bile acids in the rat. J Clin Invest. 1988 May;81(5):1593–1599. [PubMed]
  • Takikawa H, Sano N, Narita T, Uchida Y, Yamanaka M, Horie T, Mikami T, Tagaya O. Biliary excretion of bile acid conjugates in a hyperbilirubinemic mutant Sprague-Dawley rat. Hepatology. 1991 Aug;14(2):352–360. [PubMed]
  • Kartenbeck J, Leuschner U, Mayer R, Keppler D. Absence of the canalicular isoform of the MRP gene-encoded conjugate export pump from the hepatocytes in Dubin-Johnson syndrome. Hepatology. 1996 May;23(5):1061–1066. [PubMed]
  • Nishida T, Gatmaitan Z, Roy-Chowdhry J, Arias IM. Two distinct mechanisms for bilirubin glucuronide transport by rat bile canalicular membrane vesicles. Demonstration of defective ATP-dependent transport in rats (TR-) with inherited conjugated hyperbilirubinemia. J Clin Invest. 1992 Nov;90(5):2130–2135. [PubMed]
  • Huber M, Guhlmann A, Jansen PL, Keppler D. Hereditary defect of hepatobiliary cysteinyl leukotriene elimination in mutant rats with defective hepatic anion excretion. Hepatology. 1987 Mar–Apr;7(2):224–228. [PubMed]
  • Cole SP, Bhardwaj G, Gerlach JH, Mackie JE, Grant CE, Almquist KC, Stewart AJ, Kurz EU, Duncan AM, Deeley RG. Overexpression of a transporter gene in a multidrug-resistant human lung cancer cell line. Science. 1992 Dec 4;258(5088):1650–1654. [PubMed]
  • Jedlitschky G, Leier I, Buchholz U, Center M, Keppler D. ATP-dependent transport of glutathione S-conjugates by the multidrug resistance-associated protein. Cancer Res. 1994 Sep 15;54(18):4833–4836. [PubMed]
  • Leier I, Jedlitschky G, Buchholz U, Cole SP, Deeley RG, Keppler D. The MRP gene encodes an ATP-dependent export pump for leukotriene C4 and structurally related conjugates. J Biol Chem. 1994 Nov 11;269(45):27807–27810. [PubMed]
  • Müller M, Meijer C, Zaman GJ, Borst P, Scheper RJ, Mulder NH, de Vries EG, Jansen PL. Overexpression of the gene encoding the multidrug resistance-associated protein results in increased ATP-dependent glutathione S-conjugate transport. Proc Natl Acad Sci U S A. 1994 Dec 20;91(26):13033–13037. [PubMed]
  • Jedlitschky G, Leier I, Buchholz U, Barnouin K, Kurz G, Keppler D. Transport of glutathione, glucuronate, and sulfate conjugates by the MRP gene-encoded conjugate export pump. Cancer Res. 1996 Mar 1;56(5):988–994. [PubMed]
  • Thomas GA, Barrand MA, Stewart S, Rabbitts PH, Williams ED, Twentyman PR. Expression of the multidrug resistance-associated protein (MRP) gene in human lung tumours and normal tissue as determined by in situ hybridisation. Eur J Cancer. 1994;30A(11):1705–1709. [PubMed]
  • Flens MJ, Zaman GJ, van der Valk P, Izquierdo MA, Schroeijers AB, Scheffer GL, van der Groep P, de Haas M, Meijer CJ, Scheper RJ. Tissue distribution of the multidrug resistance protein. Am J Pathol. 1996 Apr;148(4):1237–1247. [PubMed]
  • Paulusma CC, Bosma PJ, Zaman GJ, Bakker CT, Otter M, Scheffer GL, Scheper RJ, Borst P, Oude Elferink RP. Congenital jaundice in rats with a mutation in a multidrug resistance-associated protein gene. Science. 1996 Feb 23;271(5252):1126–1128. [PubMed]
  • Büchler M, König J, Brom M, Kartenbeck J, Spring H, Horie T, Keppler D. cDNA cloning of the hepatocyte canalicular isoform of the multidrug resistance protein, cMrp, reveals a novel conjugate export pump deficient in hyperbilirubinemic mutant rats. J Biol Chem. 1996 Jun 21;271(25):15091–15098. [PubMed]
  • Keppler D, Konig J. Hepatic canalicular membrane 5: Expression and localization of the conjugate export pump encoded by the MRP2 (cMRP/cMOAT) gene in liver. FASEB J. 1997 Jun;11(7):509–516. [PubMed]
  • Taniguchi K, Wada M, Kohno K, Nakamura T, Kawabe T, Kawakami M, Kagotani K, Okumura K, Akiyama S, Kuwano M. A human canalicular multispecific organic anion transporter (cMOAT) gene is overexpressed in cisplatin-resistant human cancer cell lines with decreased drug accumulation. Cancer Res. 1996 Sep 15;56(18):4124–4129. [PubMed]
  • Krishnamachary N, Ma L, Zheng L, Safa AR, Center MS. Analysis of MRP gene expression and function in HL60 cells isolated for resistance to adriamycin. Oncol Res. 1994;6(3):119–127. [PubMed]
  • Hipfner DR, Gauldie SD, Deeley RG, Cole SP. Detection of the M(r) 190,000 multidrug resistance protein, MRP, with monoclonal antibodies. Cancer Res. 1994 Nov 15;54(22):5788–5792. [PubMed]
  • Grant CE, Valdimarsson G, Hipfner DR, Almquist KC, Cole SP, Deeley RG. Overexpression of multidrug resistance-associated protein (MRP) increases resistance to natural product drugs. Cancer Res. 1994 Jan 15;54(2):357–361. [PubMed]
  • Schaub T, Ishikawa T, Keppler D. ATP-dependent leukotriene export from mastocytoma cells. FEBS Lett. 1991 Feb 11;279(1):83–86. [PubMed]
  • Leier I, Jedlitschky G, Buchholz U, Keppler D. Characterization of the ATP-dependent leukotriene C4 export carrier in mastocytoma cells. Eur J Biochem. 1994 Mar 1;220(2):599–606. [PubMed]
  • Böhme M, Müller M, Leier I, Jedlitschky G, Keppler D. Cholestasis caused by inhibition of the adenosine triphosphate-dependent bile salt transport in rat liver. Gastroenterology. 1994 Jul;107(1):255–265. [PubMed]
  • Towbin H, Staehelin T, Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. [PubMed]
  • Senafi SB, Clarke DJ, Burchell B. Investigation of the substrate specificity of a cloned expressed human bilirubin UDP-glucuronosyltransferase: UDP-sugar specificity and involvement in steroid and xenobiotic glucuronidation. Biochem J. 1994 Oct 1;303(Pt 1):233–240. [PubMed]
  • Fenn JB, Mann M, Meng CK, Wong SF, Whitehouse CM. Electrospray ionization for mass spectrometry of large biomolecules. Science. 1989 Oct 6;246(4926):64–71. [PubMed]
  • Böhme M, Büchler M, Müller M, Keppler D. Differential inhibition by cyclosporins of primary-active ATP-dependent transporters in the hepatocyte canalicular membrane. FEBS Lett. 1993 Oct 25;333(1-2):193–196. [PubMed]
  • Adachi Y, Kobayashi H, Kurumi Y, Shouji M, Kitano M, Yamamoto T. ATP-dependent taurocholate transport by rat liver canalicular membrane vesicles. Hepatology. 1991 Oct;14(4 Pt 1):655–659. [PubMed]
  • Müller M, Ishikawa T, Berger U, Klünemann C, Lucka L, Schreyer A, Kannicht C, Reutter W, Kurz G, Keppler D. ATP-dependent transport of taurocholate across the hepatocyte canalicular membrane mediated by a 110-kDa glycoprotein binding ATP and bile salt. J Biol Chem. 1991 Oct 5;266(28):18920–18926. [PubMed]
  • Nishida T, Gatmaitan Z, Che M, Arias IM. Rat liver canalicular membrane vesicles contain an ATP-dependent bile acid transport system. Proc Natl Acad Sci U S A. 1991 Aug 1;88(15):6590–6594. [PubMed]
  • Loe DW, Almquist KC, Cole SP, Deeley RG. ATP-dependent 17 beta-estradiol 17-(beta-D-glucuronide) transport by multidrug resistance protein (MRP). Inhibition by cholestatic steroids. J Biol Chem. 1996 Apr 19;271(16):9683–9689. [PubMed]
  • Guhlmann A, Krauss K, Oberdorfer F, Siegel T, Scheuber PH, Müller J, Csuk-Glänzer B, Ziegler S, Ostertag H, Keppler D. Noninvasive assessment of hepatobiliary and renal elimination of cysteinyl leukotrienes by positron emission tomography. Hepatology. 1995 Jun;21(6):1568–1575. [PubMed]
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